The Bangiales represent an ancient lineage within red algae that are characterized by a life history featuring a special transitional stage from diploid to haploid known as the conchosporangia stage. However, the regulatory mechanisms governing the initiation of this stage by changes in environmental conditions are not well understood. This study analyzed the changes in phytohormones and H2O2 content during the development of conchosporangia. It also compared the gene expression changes in the early development of conchosporangia through transcriptome analysis. The findings revealed that H2O2 was shown to be the key signal initiating the transition from conchocelis to conchosporangia in Pyropia haitanensis. Phytohormone analysis showed a significant increase in 1-aminocylopropane-1-carboxylic acid (ACC) levels during conchosporangia maturation, while changes in environmental conditions were found to promote the rapid release of H2O2. H2O2 induction led to conchosporangia development, and ACC enhanced both H2O2 production and conchosporangia development. This promotive effect was inhibited by the NADPH oxidase inhibitor diphenylene iodonium and the H2O2 scavenger N, N\'-dimethylthiourea. The balance of oxidative-antioxidative mechanisms was maintained by regulating the activities and transcriptional levels of enzymes involved in H2O2 production and scavenging. Transcriptome analysis in conjunction with evaluation of enzyme and transcription level changes revealed upregulation of protein and sugar synthesis along with modulation of energy supply under the conditions that induced maturation, and exogenous ACC was found to enhance the entire process. Overall, this study demonstrates that ACC enhances H2O2 promotion of the life cycle switch responsible for the transition from a vegetative conchocelis to a meiosis-preceding conchosporangia stage in Bangiales species.

Pyropia is an economically important edible red alga worldwide. The aquaculture industry and Pyropia production have grown considerably in recent decades. Microbial communities inhabit the algal surface and produce a variety of compounds that can influence host adaptation. Previous studies on the Pyropia microbiome were focused on the microbial components or the function of specific microbial lineages, which frequently exclude metabolic information and contained only a small fraction of the overall community. Here, we performed a genome-centric analysis to study the metabolic potential of the Pyropia haitanensis phycosphere bacteria. We reconstructed 202 unique metagenome-assembled genomes (MAGs) comprising all major taxa present within the P. haitanensis microbiome. The addition of MAGs to the genome tree containing all publicly available Pyropia-associated microorganisms increased the phylogenetic diversity by 50% within the bacteria. Metabolic reconstruction of the MAGs showed functional redundancy across taxa for pathways including nitrate reduction, taurine metabolism, organophosphorus, and 1-aminocyclopropane-1-carboxylate degradation, auxin, and vitamin B12 synthesis. Some microbial functions, such as auxin and vitamin B12 synthesis, that were previously assigned to a few Pyropia-associated microorganisms were distributed across the diverse epiphytic taxa. Other metabolic pathways, such as ammonia oxidation, denitrification, and sulfide oxidation, were confined to specific keystone taxa.

Conchosporangia maturation is crucial for the yield of Pyropia/Porphyra. However, the molecular mechanisms underlying this process are poorly understood. In this study, we selected two strains of Pyropia haitanensis that show significant differences in conchosporangia maturation as materials to produce RNA-Seq libraries. Then, we identified key molecular pathways and genes involved in conchosporangia maturation by conducting a weighted gene co-expression network analysis. Two specific modules were identified, and included functions such as phosphorus metabolism, lipid metabolism, and the phosphatidylinositol signaling system. The hub genes that responded positively during conchosporangia maturation encoded diacylglycerol kinase (DGK) and phosphatidylinositol-3-phosphate-5-kinase, which are involved in the synthesis of phosphatidic acid, a key component of lipid metabolism. A full-length DGK sequence of P. haitanensis, designated as PhDGK1, was obtained by rapid-amplification of cDNA ends. Conserved motif and phylogenetic tree analyses showed that PhDGK1 belongs to DGK Cluster II. The transcript level of PhDGK1 increased during conchosporangia maturation in both strains, but increased earlier, and to higher levels, in the early-maturing strain than in the late-maturing strain. This pattern of gene expression was consistent with the patterns of maturity and changes in pigment contents. These results indicate that lipid metabolism plays a key role in regulating conchosporangia maturation in Pyropia spp., and that PhDGK1 might be a useful molecular marker for breeding new early-maturing strains.

Up to now, complicated organoarsenicals were mainly identified in marine organisms, suggesting that these organisms play a critical role in arsenic biogeochemical cycling because of low phosphate and relatively high arsenic concentration in the marine environment. However, the response of marine macroalgae to inorganic arsenic remains unknown. In this study, Pyropia haitanensis were exposed to arsenate [As(V)] (0.1, 1, 10, 100 μM) or arsenite [As(III)] (0.1, 1, 10 μM) under laboratory conditions for 3 d. The species of water-soluble arsenic, the total concentration of lipid-soluble and cell residue arsenic of the algae cells was analyzed. As(V) was mainly transformed into oxo-arsenosugar-phosphate, with other arsenic compounds such as monomethylated, As(III), demethylated arsenic and oxo-arsenosugar-glycerol being likely the intermediates of arsenosugar synthesis. When high concentration of As(III) was toxic to P. haitanensis, As(III) entered into the cells and was transformed into less toxic organoarsenicals and As(V). Transcriptome results showed genes involved in DNA replication, mismatch repair, base excision repair, and nucleotide excision repair were up-regulated in the algae cells exposed to 10 μM As(V), and multiple genes involved in glutathione metabolism and photosynthetic were up-regulated by 1 μM As(III). A large number of ABC transporters were down-regulated by As(V) while ten genes related to ABC transporters were up-regulated by As(III), indicating that ABC transporters were involved in transporting As(III) to vacuoles in algae cells. These results indicated that P. haitanensis detoxifies inorganic arsenic via transforming them into organoarsenicals and enhancing the isolation of highly toxic As(III) in vacuoles.

The blooms of harmful microalgae represent a prominent threat to fisheries, public health, and economies throughout the world. Recent studies have shown that certain macroalgae release allelochemicals that can inhibit the growth of bloom-forming microalgae. In this study, we found that the macroalga Pyropia haitanensis significantly inhibited growth of the harmful bloom-forming microalgae Pseudo-nitzschia pungens and Pseudo-nitzschia multiseries. The inhibitory-effect of the live thali of P. haitanensis was highest, followed by that of dry powder, water-soluble extract, and culture medium filtrate. The Pseudo-nitzschia species died 96 h after exposure to 5-10 g fresh-weight L-1 of P. haitanensis live thalli. Furthermore, an aqueous extract of P. haitanensis suppressed the growth of P. pungens and P. multiseries, thereby indicating that P. haitanensis contains stable allelopathic substances that cause the observed inhibitory-effects. On the basis of these findings, we conclude that the macroalga P. haitanensis would have potential utility in controlling the blooms of Pseudo-nitzschia species.

A sulfated galactan LP-G2 was isolated and purified from red alga, Pyropia haitanensis. It was a branched polysaccharide with average molecular weight about 8381 Da, composed of Gal, Glc, GalA and Ara. The structure of LP-G2 was determined using IR and NMR spectroscopy. It was composed of →4)-β-d-galactose→4)-α-l-galactose-6- sulfate segments, with β-d-Glc and α-d-galactose unit substituted at the 6-position of α-l-galactose. Functional analysis showed that LP-G2 inhibited complement activation on both the classic and alternative pathways with CH50 value of 3.08 ± 0.25 mg/mL and AP50 value of 2.23 ± 0.20 mg/mL, respectively. Preliminary mechanism studies by using complement component depleted-sera indicated that LP-G2 selectively interacts with C1q, C2, C4 and C9. The results suggested that LP-G2 could be of potential benefits in treatment of the complement associated diseases.

An antioxidant polysaccharide, porphyran, from red algae Pyropia haitanensis, is introduced as a protective agent against neurotoxicity-induced amyloid β peptide (Aβ) of Alzheimer\'s disease (AD) mice. Then the activity of acetylcholinesterase (AChE) and choline acetyltransferase (CHAT) in the cortical and hippocampal tissue were examined by colorimetric method. Results showed that porphyran significantly ameliorated the learning and memory impairment induced by Aβ1-40. Biochemical analysis showed that porphyran increased ChAT activity and decreased AChE activity in the cortical and hippocampal tissue. The mechanism may be related with the increase of cerebral acetylcholine content. Porphyran has a potential of developing anti-aging drug.

BACKGROUND: Pyropia haitanensis, distributes in the intertidal zone, can tolerate water losses exceeding 90%. However, the mechanisms enabling P. haitanensis to survive harsh conditions remain uncharacterized. To elucidate the mechanism underlying P. haitanensis desiccation tolerance, we completed an integrated analysis of its transcriptome and proteome as well as transgenic Chlamydomonas reinhardtii carrying a P. haitanensis gene.
RESULTS: P. haitanensis rapidly adjusted its physiological activities to compensate for water losses up to 60%, after which, photosynthesis, antioxidant systems, chaperones, and cytoskeleton were activated to response to severe desiccation stress. The integrative analysis suggested that transketolase (TKL) was affected by all desiccation treatments. Transgenic C. reinhardtii cells overexpressed PhTKL grew better than the wild-type cells in response to osmotic stress.
CONCLUSIONS: P. haitanensis quickly establishes acclimatory homeostasis regarding its transcriptome and proteome to ensure its thalli can recover after being rehydrated. Additionally, PhTKL is vital for P. haitanensis desiccation tolerance. The present data may provide new insights for the breeding of algae and plants exhibiting enhanced desiccation tolerance.

BACKGROUND: The heteroside floridoside is a primary photosynthetic product that is known to contribute to osmotic acclimation in almost all orders of Rhodophyta. However, the encoding genes and enzymes responsible for the synthesis of floridoside and its isomeric form, L- or D-isofloridoside, are poorly studied.
RESULTS: Here, four putative trehalose-6-phosphate synthase (TPS) genes, designated as PhTPS1, PhTPS2, PhTPS3, and PhTPS4, were cloned and characterized from the red alga Pyropia haitanensis (Bangiophyceae). The deduced amino acid sequence is similar to the annotated TPS proteins of other organisms, especially the UDP-galactose substrate binding sites of PhTPS1, 2, which are highly conserved. Of these, PhTPS1, 4 are involved in the biosynthesis of floridoside and isofloridoside, with isofloridoside being the main product. PhTPS3 is an isofloridoside phosphate synthase, while PhTPS2 exhibits no activity. When challenged by desiccation, high temperature, and salt stress, PhTPS members were expressed to different degrees, but the responses to thermal stress and desiccation were stronger.
CONCLUSIONS: Thus, in P. haitanensis, PhTPSs encode the enzymatical activity of floridoside and isofloridoside phosphate synthase and are crucial for the abiotic stress defense response.

Intertidal macroalgae are highly resistant to hypersaline stress conditions. However, the underlying mechanism remains unknown. In the present study, the mechanism behind Pyropia haitanensis responses to two hypersaline stress conditions [100‰ (HSS_100) and 110‰ (HSS_110)] was investigated via analyses of physiological and transcriptomic changes. We observed that the differences between the responses of Py. haitanensis to HSS_100 and HSS_110 conditions involved the following three aspects: osmotic regulation, ionic homeostasis, and adjustment to secondary stresses. First, the water retention of Py. haitanensis was maintained through increased expansin production under HSS_100 conditions, while cell wall pectin needed to be protected from hydrolysis via the increased abundance of a pectin methylesterase inhibitor under HSS_110 conditions. Meanwhile, Py. haitanensis achieved stable and rapid osmotic adjustments because of the coordinated accumulation of inorganic ions (K+, Na+, and Cl-) and organic osmolytes (glycine betaine and trehalose) under HSS_100 conditions, but not under HSS_110 conditions. Second, Py. haitanensis maintained a higher K+/Na+ ratio under HSS_100 conditions than under HSS_110 conditions, mainly via the export of Na+ into the apoplast rather than compartmentalizing it into the vacuoles, and the enhanced uptake and retention of K+. However, K+/Na+ homeostasis was not completely disrupted during a short-term exposure to HSS_110 conditions. Finally, the Py. haitanensis antioxidant system scavenged more ROS and synthesized more heat shock proteins under HSS_100 conditions than under HSS_110 conditions, although thalli may have been able to maintain a certain redox balance during a short-term exposure to HSS_110 conditions. These differences may explain why Py. haitanensis can adapt to HSS_100 conditions rather than HSS_110 conditions, and also why the thalli exposed to HSS_110 conditions can recover after being transferred to normal seawater. Thus, the data presented herein may elucidate the mechanisms enabling Pyropia species to tolerate the sudden and periodic changes in salinity typical of intertidal systems.